| Assay Method Information | |
| | Inhibitory Activity Assay |
| Description: | Capillary electrophoresis was used to detect the phosphorylation conversion rate of the substrate peptide to determine the IC50 value of the test compound for inhibiting the kinase (ABL1-WT). The maximum concentration of the compound tested in the test was 1000 nM, with 3-fold dilution, and a total of 12 concentrations (1000-0.0056 nM). First, an enzyme reaction system (the concentration of enzyme ABL1-WT was 1.3 nM, the concentration of substrate FLPeptide2 was 1.5 μM, and the reaction factor was 10 mM MgCl2) was prepared. After incubation at room temperature for 30 min, 5 L of 4×ATP solution was added to start the enzyme reaction. After reacting at room temperature for 90 min, the reaction was stopped by adding a stop buffer (containing 0.5 M EDTA). The sample was analyzed using an EZ reader (analysis conditions: pressure −1.5 PSI, upper voltage current −2250V, lower voltage current −500V, separation time 40 sec, and system delay 100.0 sec). |
| Affinity data for this assay | |
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